Parasite 25, 19 (2018) © M.J. Walker et al., published by EDP Sciences, 2018 https://doi.org/10.1051/parasite/2018017 Available online at: www.parasite-journal.org RESEARCH ARTICLE A tripartite survey of hyperparasitic fungi associated with ectoparasitic flies on bats (Mammalia: Chiroptera) in a neotropical cloud forest in Panama Melissa J. Walker1,2,a, Annabel Dorrestein2,3,a, Jasmin J. Camacho4, Lauren A. Meckler5, Kirk A. Silas6, Thomas Hiller7, and Danny Haelewaters4,8,9,* 1 Department of Ecology, Environment and Evolution, La Trobe Institute for Molecular Science Bundoora, Victoria 3083, Australia 2 Current address: Hawkesbury Institute for the Environment, Western Sydney University, Science Road, Richmond, New South Wales 2753, Australia 3 Department of Animal Ecology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands 4 Department of Organismic and Evolutionary Biology, Harvard University, 26 Oxford Street, Cambridge, Massachusetts 02138, USA 5 Department of Psychological and Brain Sciences, University of Delaware, 105 The Green, Newark, Delaware 19716, USA 6 Painted Sky Road, Reading, Pennsylvania 19606, USA 7 Institute of Evolutionary Ecology and Conservation Genomics, University of Ulm, Albert-Einstein Allee 11, 89081 Ulm, Germany 8 Smithsonian Tropical Research Institute, Apartado Postal 0843-03092, Balboa, Panama 9 Herbario UCH, Universidad Autónoma de Chiriquí, Apartado Postal 0427, David, Panama Received 17 November 2017, Accepted 5 March 2018, Published online 10 April 2018 Abstract- -The Darién province in eastern Panama is one of the most unexplored and biodiverse regions in the world. The Chucantí Nature Reserve, in Serranía de Majé, consists of a diverse tropical cloud forest ecosystem. The aim of this research was to explore and study host associations of a tripartite system of bats, ectoparasitic flies on bats (Diptera, Streblidae), and ectoparasitic fungi (Ascomycota, Laboulbeniales) that use bat flies as hosts. We captured bats at Chucantí, screened each bat for presence of bat flies, and screened collected bat flies for presence of Laboulbeniales. We mistnetted for 68 mistnet hours and captured 227 bats representing 17 species. We captured Micronycteris schmidtorum, a species previously unreported in Darién. In addition, we encountered the rarely collected Platyrrhinus dorsalis, representing the westernmost report for this species. Of all captured bats, 148 carried bat flies (65%). The number of sampled bat flies was 437, representing 16 species. One species represents a new country record (Trichobius anducei) and five species represent first reports for Darién (Basilia anceps, Anatrichobius scorzai, Nycterophilia parnelli, T. johnsonae, T. parasiticus). All 74 bat fly species currently reported in Panama are presented in tabulated form. Of all screened bat flies, 30 bore Laboulbeniales fungi (7%). Based on both morphology and large ribosomal subunit (LSU) sequence data, we delimited 7 species of Laboulbeniales: Gloeandromyces nycteribiidarum (newly reported for Panama), G. pageanus, G. streblae, Nycteromyces streblidinus, and 3 undescribed species. Of the 30 infected flies, 21 were Trichobius joblingi. This species was the only host on which we observed double infections of Laboulbeniales. Keywords: Bat flies, Diptera, Ectoparasites, Hyperparasites, Laboulbeniales, Sequence-based identification of fungi Résumé- - Étude tripartite des champignons hyperparasites associés aux mouches ectoparasites des chauves-souris (Mammalia, Chiroptera) dans une forêt néotropicale au Panama. La province de Darién dans l’est du Panama est l’une des régions les plus inexplorées et les plus riches en biodiversité au monde. La réserve naturelle du Chucantí, à Serranía de Majé, est constituée d’un écosystème forestier tropical varié. Le but de cette recherche était d’explorer et d’étudier les associations hôtes d’un système tripartite de chauves-souris, de mouches ectoparasites sur chauves-souris (Diptera, Streblidae) et de *Corresponding author: [email protected] a These authors contributed equally to the manuscript This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 2 M.J. Walker et al.: Parasite 2018, 25,19 champignons ectoparasites (Ascomycota, Laboulbeniales) utilisant des diptères de chauves-souris comme hôtes. Nous avons capturé des chauves-souris à Chucantí, criblé chaque chauve-souris pour la présence de diptères, et criblé les diptères recueillis pour la présence de Laboulbeniales. Nous avons utilisé des filets japonais pendant 68 heures et avons capturé 227 chauves-souris représentant 17 espèces. Nous avons capturé Micronycteris schmidtorum, une espèce précédemment non rapportée de Darién. En outre, nous avons rencontré l’espèce rarement collectée Platyrrhinus dorsalis, ce qui représente la mention la plus occidentale de cette espèce. De toutes les chauves-souris capturées, 148 portaient des diptères (65%). Le nombre de diptères échantillonnés était de 437, représentant 16 espèces. Une espèce représente une nouvelle mention pour ce pays (Trichobius anducei)etcinq espèces représentent les premières mentions pour Darién (Basilia anceps, Anatrichobius scorzai, Nycterophilia parnelli, T. johnsonae, T. parasiticus). Les 74 espèces de diptères de chauves-souris actuellement signalées au Panama sont présentées sous forme de tableau. Parmi tous les diptères examinés, 30 portaient des champignons Laboulbeniales (7%). Sur la base de la morphologie et des données des séquences d’ADNr LSU, nous avons délimité 7 espèces de Laboulbeniales : Gloeandromyces nycteribiidarum (nouvelle mention pour Panama), G. pageanus, G. streblae, Nycteromyces streblidinus et 3 espèces non décrites. Sur les 30 mouches infectées, 21 étaient des Trichobius joblingi. Cette espèce est le seul hôte sur lequel nous avons observé des doubles infections de Laboulbeniales. Introduction much of Chucantí is recovering. This constructive reestablishment of the natural ecosystem, in conjunction Panamanian forests are under major risk of deforesta- with researchers obtaining supported access to the tion, threatening the associated biodiversity. The 2015 mountain, has spurred greater insight into the environ- Global Forest Resources Assessment reports that 62.1% of ment of this area. Accordingly, we documented bat Panama is forested; however, there was an annual change species richness and abundance at three altitudes in rate of À0.4% between 1990 and 2015 [23]. Recent efforts Chucantí. We screened all captured bats for the presence by various independent bodies and organizations have of bat flies, which in turn were screened for the presence focused on securing existing forests, reforesting farm of Laboulbeniales fungi, with the aim of reporting lands, and reporting on unidentified and known species biodiversity at different trophic levels (host–parasite– endemic to the region. In recent years, many new species parasite) and studying host associations. Below, we of plants, animals, and fungi have been reported in briefly introduce the three levels of this hyperparasitic Panamanian forests. A Google Scholar search (on study system. September 23, 2017) using the keywords “new species” and “Panama” resulted in 1,740 hits since 2017 alone. This Bats (Mammalia: Chiroptera) collaborative management has benefited Panama’s native flora and fauna, while promoting the importance of Bats are the only mammals capable of sustained flight. rainforest conservation [61,81,82,96,97]. Although re- They use echolocation and have a wide assortment of food search efforts are steadfast, difficult terrain and political sources, thereby providing important ecosystem services strife leaves many Panamanian locations isolated and like insect predation and seed dispersal [54]. The New difficult to explore. World leaf-nosed bats (family Phyllostomidae) are the One such location is the Darién province, in eastern most morphologically and ecologically diverse of all bats. Panama. Darién, host to the Darién National Park, is They have evolved extraordinarily diverse faces, skulls, one of the most pristine habitats in Central America, and and teeth, adapted to many different food types, including one of the most endemically biodiverse zones in the insects, other vertebrates, blood, fruit, and nectar [26]. world. Dividing the Panamá and Darién provinces is the Bats also differ greatly in their roosting habits; roosts vary Serranía de Majé, a 60 km long, 404 ha mountain range from more permanent and enclosed structures (caves, rock with elevations from 600 to 1480 m a.s.l. [2,71]. The crevices, mines) to ephemeral and exposed structures (leaf summit of Serranía de Majé is Cerro Chucantí, a diverse tents, plant foliage) [53]. tropical cloud forest ecosystem [71] and host to the There are around 1,200 species of bats worldwide [9]. In Chucantí Nature Reserve. However, the rough terrain, Panama, a total of 118 bat species are documented, proximity to Colombian border, and isolation of Cerro therefore representing the most diverse mammal group in Chucantí has thus far resulted in many species going the country [78]. Although these species reports are undocumented. numerous, many of the results come from lowland Prior to private purchase, parts of Chucantí Nature research, leaving many highland Panamanian regions Reserve (hereafter: Chucantí) were subject to logging, without